骨髓间充质干细胞移植对视网膜病变新生大鼠视网膜细胞凋亡的影响

Effects of bone marrow mesenchymal stem cell transplantation on retinal cell apoptosis in premature rats with retinopathy

摘要:

目的：探讨早产儿视网膜病变（ROP）大鼠玻璃体内注射骨髓间充质干细胞（BMSCs）对视网膜细胞凋亡及视网膜中神经营养素-3（NT-3）、睫状神经营养因子（CNTF）表达的影响。方法：40只7 日龄 Sprague-Dawley新生大鼠随机分为正常对照组、ROP模型组、骨髓间充质干细胞（BMSCs）移植组与磷酸缓冲液组（PBS组），每组10只。采用高氧法制成 ROP 模型，持续高氧5 d后分别给予BMSCs移植组与PBS组玻璃体内注射BMSCs和PBS。移植7 d后，采用 TUNEL/DAPI、NT-3/DAPI、CNTF/DAPI免疫荧光双标法观察BMSCs移植对视网膜细胞凋亡及NT-3、CNTF表达的影响。结果：移植7 d后，正常对照组视网膜几乎未见TUNEL+ DAPI+细胞，BMSCs移植组视网膜可见少量TUNEL+ DAPI+细胞，显著低于ROP模型组与PBS组（P0.05）；正常对照组视网膜可见极少量的NT-3+DAPI+与CNTF+DAPI+ 细胞，ROP模型组与PBS组大鼠视网膜可见少量NT-3+DAPI+与CNTF+DAPI+ 细胞，较正常对照组增多，但差异无统计学意义（P>0.05）；BMSCs移植组大鼠视网膜NT-3+DAPI+与CNTF+DAPI+ 细胞数均显著多于ROP模型组与PBS组(P0.05）。结论：BMSCs移植可减轻ROP大鼠视网膜细胞的凋亡，其机制可能与其促进ROP大鼠视网膜细胞NT-3与CNTF的表达有关。

Abstract:

OBJECTIVE: To explore the effects of marrow mesenchymal stem cell (BMSC) transplantation on retinal cells apoptosis and changes to neurotrophin-3 (NT-3 and ciliary neurotrophic factor (CNTF) in rats with retinopathy of prematurity (ROP). METHODS: Seven-day-old Sprague-Dawley rats were randomly divided into normal control (CON), ROP, BMSC transplantation (BMSCs were transplanted 5 days after oxygen conditioning) and phosphate buffered saline (PBS) groups. The ROP model was prepared according to the classic hyperoxygen method. Seven days after transplantation, TUNEL/DAPI, NT-3/API and CNTF/DAPI double-labeled immunofluorescence were used to examine the effects of BMSC transplantation on both the apoptosis of retinal cells and the expression of NT-3 and CNTF protein in the retinal cells of the ROP rats. RESULTS: Seven days after BMSC transplantation, there were few TUNEL+ DAPI+ cells observed in the CON group. There were fewer TUNEL+DAPI+ cells observed in the BMSC group than in the ROP group (P0.05). There were few NT-3+DAPI+ cells and CNTF+DAPI+ cells in the CON group. There were more NT-3+DAPI+ and CNTF+DAPI+ cells in the ROP group than in the CON group, but there was no significant difference between the ROP and CON groups (P>0.05). More NT-3+DAPI+ and CNTF+DAPI+ cells were observed in the BMSC group compared with the ROP group (P0.05). CONCLUSIONS: BMSC transplantation therapy could alleviate the apoptosis of retinal cells in ROP rats, and its mechanisms might be associated with promoting the expression of NT-3 and CNTF protein in retinal cells.